海带热休克蛋白70基因体外原核表达研究

Study on Prokaryotic Expression of Laminaria japonica Heat Shock Protein 70

在前期研究克隆得到全长海带HSP70基因的基础上,为进一步研究藻类HSP70的生物学功能,将海带HSP70基因的开放阅读框区域克隆到表达载体pEASY-E2中,并转化到大肠杆菌BL21(DE3)pLysS。将阳性重组子培养于含有AMP(100 U/mL)的LB培养基,IPTG诱导表达,SDS-PAGE电泳鉴定。经5 h诱导,其表达量达到平台期,继续培养HSP70表达量并不显著增高。5 mM IPTG诱导海带HSP70蛋白表达量高于1 mM IPTG诱导蛋白表达量。

On the early stage work of cloning full length cytosolic Laminaria japonica heat shock protein 70（LJHSP70）,to further study the biological function of seaweed HSP70 in vivo and in vitro,the open reading frame of LJHSP70 gene was subcloned into expression vector of pEASY-E2.The recombinant plasmid was transformed to E.coli BL21（DE3）pLysS.The positive recombinant was cultured in LB media with 100 U/mL ampicillin,induced by IPTG and determined by SDS-PAGE.The result showed that the expression level of LJHSP70 reached high level after 5 h of induction,after that the expression level increased very slowly.The expression level of LJHSP70 at 5 mM IPTG was higher than that at 1 mM.